TY - JOUR
T1 - Tissue factor activity of small and large extracellular vesicles in different diseases
AU - Sachetto, Ana T. A.
AU - Archibald, Sierra J.
AU - Hisada, Yohei
AU - Rosell, Axel
AU - Havervall, Sebastian
AU - van Es, Nick
AU - Nieuwland, Rienk
AU - Campbell, Robert A.
AU - Middleton, Elizabeth A.
AU - Rondina, Matthew T.
AU - Thålin, Charlotte
AU - Mackman, Nigel
N1 - Funding Information: This work was supported by the NIH NHLBI R35HL155657 (N.M.) and the John C. Parker professorship (N.M.). Funding Information: We would like to acknowledge Dr. S. Grover for helpful comments and Dr. M. Sokolsky for help with the NTA. In addition, the authors would like to thank the healthy donors and patients that donated blood. This work was supported by the NIH NHLBI R35HL155657 (N.M.) and the John C. Parker professorship (N.M.). Healthy individuals and patients provided informed consent. The study's protocols were approved by: the Institutional Review Board of the University of North Carolina at Chapel Hill (protocol 14-2108); the Institutional Review Board of the University of Utah (IRB number 00102638); the institutional review board of the hospital and registered in ClinicalTrials.gov (identification number: NCT02095925); the Swedish Ethical Review Board (COMMUNITY study [COVID-19 Biomarker and Immunity] dnr 2020-01653). A.T.A.S. and N.M. designed experiments, interpreted data, and edited the manuscript. A.T.A.S. and S.A. conducted experiments, analyzed data, and wrote the manuscript. R.A.C. E.A.M. and M.T.R. provided the plasma samples from patients with sepsis and edited the manuscript. N.v.E. and R.N. provided the plasma samples from patients with pancreatic cancer and edited the manuscript. A.R. S.H. and C.T. provided the plasma samples from patients with COVID-19 and edited the manuscript. Y.H. edited the manuscript. There are no competing interests to disclose. Publisher Copyright: © 2023 The Author(s)
PY - 2023/3/1
Y1 - 2023/3/1
N2 - Background: Circulating procoagulant extracellular vesicles (EVs) are increased in diseases, such as cancer, sepsis, and COVID-19. EV tissue factor (TF) activity is associated with disseminated intravascular coagulation in sepsis and venous thrombosis in patients with pancreatic cancer and COVID-19. EVs are commonly isolated by centrifugation at ∼20,000 g. Objectives: In this study, we analyzed the TF activity of 2 EV populations enriched for large and small EVs in patients with either sepsis, pancreatic cancer, or COVID-19. Methods: EVs were isolated from plasma by sequential centrifugation at 20,000 g (large EVs, LEVs) and then 100,000 g (small EVs, SEVs). We analyzed EVs from plasma prepared from whole blood samples from healthy individuals with or without lipopolysaccharide (LPS) stimulation as well as EVs from plasma samples from patients with either sepsis, pancreatic cancer, or COVID-19. TF-dependent (EV-TF activity) and TF-independent factor Xa (FXa) generation of the EVs was measured. Results: LPS increased EV-TF activity in LEVs but not SEVs. Similarly, in 2 patients with sepsis who had EV-TF activity above the background of the assay we observed EV-TF activity in LEVs but not SEVs. Patients with pancreatic cancer or COVID-19 had circulating EV-TF activity in both LEVs and SEVs. Conclusion: We recommend that EVs are isolated from plasma from patients by centrifugation at 100,000 g rather than 20,000 g to obtain a more accurate measure of levels of circulating EV-TF activity.
AB - Background: Circulating procoagulant extracellular vesicles (EVs) are increased in diseases, such as cancer, sepsis, and COVID-19. EV tissue factor (TF) activity is associated with disseminated intravascular coagulation in sepsis and venous thrombosis in patients with pancreatic cancer and COVID-19. EVs are commonly isolated by centrifugation at ∼20,000 g. Objectives: In this study, we analyzed the TF activity of 2 EV populations enriched for large and small EVs in patients with either sepsis, pancreatic cancer, or COVID-19. Methods: EVs were isolated from plasma by sequential centrifugation at 20,000 g (large EVs, LEVs) and then 100,000 g (small EVs, SEVs). We analyzed EVs from plasma prepared from whole blood samples from healthy individuals with or without lipopolysaccharide (LPS) stimulation as well as EVs from plasma samples from patients with either sepsis, pancreatic cancer, or COVID-19. TF-dependent (EV-TF activity) and TF-independent factor Xa (FXa) generation of the EVs was measured. Results: LPS increased EV-TF activity in LEVs but not SEVs. Similarly, in 2 patients with sepsis who had EV-TF activity above the background of the assay we observed EV-TF activity in LEVs but not SEVs. Patients with pancreatic cancer or COVID-19 had circulating EV-TF activity in both LEVs and SEVs. Conclusion: We recommend that EVs are isolated from plasma from patients by centrifugation at 100,000 g rather than 20,000 g to obtain a more accurate measure of levels of circulating EV-TF activity.
KW - COVID-19
KW - cancer
KW - extracellular vesicles
KW - procoagulant activity
KW - sepsis
KW - tissue factor
UR - http://www.scopus.com/inward/record.url?scp=85151463173&partnerID=8YFLogxK
U2 - https://doi.org/10.1016/j.rpth.2023.100124
DO - https://doi.org/10.1016/j.rpth.2023.100124
M3 - Article
C2 - 37012986
SN - 2475-0379
VL - 7
JO - Research and practice in thrombosis and haemostasis
JF - Research and practice in thrombosis and haemostasis
IS - 3
M1 - 100124
ER -